OBJECTIVETo rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis, a multiplex PCR was developed.

METHODSIn the process of this reaction, four distinct DNA targets were amplified. One target was based on the serotype 2 (and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP, EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control, and 49 clinical specimens were detected by the multiplex PCR assay.

RESULTSAll PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp+, epf+, epf*+ and sly+ were 16/48, 14/48, 12/48, 3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains.

CONCLUSIONThe results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis.

Bacterial Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Streptococcus suis ; genetics ; pathogenicity ; Virulence Factors ; genetics

BACKGROUNDOsteosarcoma is characterized by high neovascularization and a high propensity for metastasis through bloodstream. This study was to examine whether there is evidence for vasculogenic mimicry in osteosarcoma and to illustrate mechanism of tumor blood vessels formation in osteosarcoma.

METHODSOsteosarcoma cell lines (U-2OS) were tested for their ability to form tubular networks in three-dimensional culture containing type I collagen. The structures of the tubular networks were observed with phase contrast microscope and transmission electron microscope (TEM). Morphometric studies using hematoxylin and eosin (HE) stain and CD31 immunohistochemical stain to show tumor-lined channels in human osteosarcoma were also performed.

RESULTSObservation with light microscope and TEM showed that highly aggressive osteosarcoma cell lines (U-2OS) formed networks containing channels when grown in three-dimensional culture containing type I collagen, in the absence of endothelial cells or fibroblasts. Morphometric observation using HE stain and CD31 immunohistochemical stain showed that tumor cell-lined channels were also detected in vivo in osteosarcoma; by comparison, all vascular areas in the pedicle of osteochondroma or outside osteochondroma were endothelial-lined.

CONCLUSIONThese observations strongly suggest that aggressive osteosarcoma cells may generate vascular channels that facilitate tumor perfusion independent of tumor angiogenesis and have the ability of vasculogenic mimicry.

Bone Neoplasms ; pathology ; ultrastructure ; Cell Line, Tumor ; Collagen ; biosynthesis ; Humans ; Immunohistochemistry ; Neovascularization, Pathologic ; pathology ; Osteosarcoma ; pathology

OBJECTIVETo observe and assess the immunosuppressive effect of applying bailing capsule (BLC, a dry powder preparation of Cordyceps sinensis mycelia), after renal transplantation, its influence on other systems of organism, and to explore the possible therapeutic mechanism.

METHODSOne hundred and twenty-one recipients of renal homo-allograft were randomly divided into two groups. The 64 cases in Group A was treated with cyclosporin A (Cs A) + prednisone (pred) + azathioprine (Aza), the 57 in Group B treated with Cs A + pred + BLC. They were followed-up for 1-2 year by checking up blood routine, urine routine, liver and renal function, blood electrolytes, glucose and lipids, and uric acid for 2 times every week in the first month after transplantation, followed by proper re-examination of these items according to various condition.

RESULTSThere was no significant difference between the two groups in aspects of graft survival rate, occurrence of reject reaction, renal function recovery, blood electrolytes and blood glucose levels. However, as compared with Group A, in Group B, levels of urinary erythrocytes and leucocytes, blood alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol, uric acid as well as the incidence of infection were significantly lower, and blood high density lipoprotein, serum total protein, albumin, RBC and WBC count were significantly higher.

CONCLUSIONBLC could effectively prevent the reject response after renal transplantation, protect renal and liver function, stimulate hemopoietic function, improve hypoproteinemia and hyperlipidemia, reduce the infection, etc., therefore, it is an ideal immunosuppressor after organ transplantation.

Adult ; Capsules ; Cordyceps ; Cyclosporine ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis ; drug therapy ; surgery ; Graft Rejection ; prevention & control ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; Male ; Phytotherapy ; Postoperative Period ; Prednisone ; therapeutic use

Objective To evaluate the association between total fluid intake and the time of urination per day and the risk of bladder cancer. Methods A population-based case-control study was conducted in urban Shanghai, China, during January 1996 to December 1998. The study included 608 incident cases of bladder cancer and 607 age- and sex-matched controls. Unconditional logistic regression models were used to estimate the odds ratios(ORs)and their corresponding 95%confidence intervals(95%CIs)for bladder cancer associated with frequency of urination, after adjusted for age, gender, smoking status, history of occupation with high risk, history of bladder infections, body mass index and other confounding factors. The level of statistical significance was set at 0.05(two-sided). Results No significant trend was observed for the association between total fluid intake, time of nighttime urination and the risk of bladder cancer. Increasing time of urination during daytime was associated with decreased risk of bladder cancer(P for trend=0.014). ORs(95%CIs)for subjects who voided 4 times, 5 times and 6 or more times per day[0.72(0.49-1.05),0.60(0.41-0.87)and 0.62(0.43-0.90), respectively], when compared with those with less than 4times per day after adjustment of confounding factors. Data showed that smokers and nonsmokers who voided at least 6 times per day had the ORs of 0.72(95%CI: 0.45-1.15)and 0.46(95%CI:0.25-0.87)when compared to their counterparts who voided 3 times or less per day during the daytime. Subjects who urinated at least 6 times per day and consumed more than 1500 ml of total fluid per day experienced a significant 57% reduction in risk compared to subjects who urinated 3 times or less and consumed less than 750 ml of total daily fluid intake. Conclusion Increased urination frequency and total fluid intake, especially among those who never smoked might be associated with a reduced risk of bladder cancer.

Aged ; Blood Glucose ; metabolism ; Female ; Humans ; Hyperglycemia ; epidemiology ; etiology ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Postoperative Period ; Retrospective Studies

OBJECTIVETo explore the pathogenesis of tumors by blocking the normal differentiation process of stem cells.

METHODSBone marrow mesenchymal stem cells (BMSCs) from rats were isolated, cultured and purified by whole bone marrow adherence method. The rat BMSCs were induced to differentiate into adipocytes with dexamethasone, insulin and indomethacin. Blockage of the differentiation process was induced by 3-methylcholanthrene (3-MC).

RESULTSThe differentiation experiment showed that at 30 days after the induction, oil red O staining-positive cells occurred with increased intracytolasmic lipid droplets, characteristic for adipocytes. The differentiation blockage experiment showed that at 30 days after induction, the deposits of oil red O staining-cytoplasmic lipid droplets was significantly reduced, indicating that the blocked cells were adipocytes, but not fully differentiated. Morphological identification showed that cell contact inhibition disappeared, abnormal cell nuclei, increased number of micronucleus aberration and karyotype abnormalities, indicating that malignant transformation of the stem cells occurred after the differentiation blockage.

CONCLUSIONSThe results of this study show a blockage of the differentiation of that stem cells at the intermediate phase, and a tendency of malignant transformation of the stem cells. The results of our study provide new evidence that cancer stem cells may be originated by suppression of stem cell differentiation.

Adipocytes ; cytology ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cell Transformation, Neoplastic ; Cells, Cultured ; Dexamethasone ; pharmacology ; Drug Combinations ; Female ; Indomethacin ; pharmacology ; Insulin ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Methylcholanthrene ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the chronic effects of intracoronary autologous bone marrow mononuclear cell (BM-MNCs) transplantation in patients with refractory heart failure (RIHF) after myocardial infarction.

METHODSThirty patients with RIHF (LVEF < 40%) were enrolled in this nonrandomized study, autologous BM-MNCs (5.0 +/- 0.7) x 10(7) were transplanted with via infarct-related coronary artery in 16 patients and 14 patients received standard medical therapy served as control. Baseline and follow up evaluations included complete clinical evaluations, plasma BNP, ANP, ET-1 measurements, echocardiography, PET, and Holter monitoring.

RESULTSBaseline characteristics were similar between the 2 groups. There were no major periprocedural complications. One patient developed ventricular premature contractions during cell infusion for several seconds and recovered spontaneously. Compared to pre-transplantation, plasma BNP and ET-1 significantly decreased and plasma ANP significantly increased at 7 days post transplantation; 6 minutes walking distance increased from (72.1 +/- 31.5) to (201.6 +/- 23.3) m (P < 0.01), LVEF increased 9.9% (P < 0.001) and FDG-PET revealed vital myocardium area increased (10.3 +/- 3.4)% (P < 0.01) at 3 months after BM-MNCs transplantation. At 6 months follow up, the NYHA class improved from (3.4 +/- 0.1 to 2.4 +/- 0.2, P < 0.001) and no patient died and 1 patient rehospitalized due to lower extremities edema. In control group, LVEF decreased 7.2% compared to baseline (P < 0.001) and was significantly lower than transplantation group at 3 months (P < 0.001). At 6 months follow up, the NYHA class increased from (3.5 +/- 0.1 to 3.9 +/- 0.1, P < 0.05), 2 patients died and 10 patients rehospitalized due to aggravated heart failure.

CONCLUSIONPresent study demonstrates that intracoronary transplantation of autologous BM-MNCs is safe and effective for treating patients with RIHF after myocardial infarction.

Bone Marrow Transplantation ; Coronary Vessels ; surgery ; Follow-Up Studies ; Heart Failure ; complications ; Humans ; Mesenchymal Stem Cell Transplantation ; Monocytes ; transplantation ; Myocardial Infarction ; surgery ; Myocardial Ischemia ; complications ; Transplantation, Autologous

Objective: To investigate the feasibility of multiplex real-time RT-PCR with fluorescent probes in early screening of Ph-like acute lymphoblastic leukemia (ALL) and analyze the clinical feature and prognos. Method: A total of 118 adult B-ALL patients diagnosed between October 2010 and March 2016 were enrolled in this study. Multiplex RT-PCR was used to detect the Ph-like ALL related fusion gene and CRLF2 expression in 58 BCR-ABL and MLL rearrangement negative patients. The clinical features, treatment response and prognosis were analyzed in Ph-like fusion gene positive and/or CRLF2 over-expression patients. Result: Among 58 patients, 9 patients (9/58, 15.5%) showed Ph-like ALL related fusion genes positive and 10 patients (10/58, 17.2%) showed CRLF2 over-expression. There were statistical differences in age, WBC count, immunophenotypes, cytogenetics and risk stratification among Ph-like fusion gene positive or CRLF2 over-expression patients, Ph⁺ patients, MLL⁺ patients and B-other patients. The 2-year overall survival rates were 65%, 47%, 64% and 74% respectively among these four groups (P=0.043) . The 2-year relapse free survival rates were 51%, 39%, 62% and 70% respectively among these four groups (P=0.010) . Conclusion Routine screening of Ph-like ALL by multiplex RTPCR is feasible.

AIM:To investigate the effect of suberoylanilide hydroxamic acid ( SAHA) on the proliferation and apoptosis of human hepatocellular carcinoma HepG 2 cells and to explore its possible mechanism .METHODS: HepG2 cells were treated with SAHA at different concentrations for 48 h.The proliferation of HepG2 cells was detected by real-time cellular analysis.The protein levels of acetylated histones H3K9 and H3K27, glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase ( PERK ) and p-PERK were determined by Western blot .The cell apoptosis was analyzed by flow cytometry .RESULTS:Compared with control group , treatment with SAHA at 0.1μmol/L and 1 μmol/L for 48 h showed no significant inhibitory effect on the proliferation of HepG 2 cells, while SAHA at 6 μmol/L and 12 μmol/L significantly inhibited the proliferation of HepG 2 cells (P<0.05).The results of Western blot showed that the protein levels of acH3K9, acH3K27, GRP78 and p-PERK increased significantly after treated with SAHA at diffe-rent concentrations for 48 h, while the protein level of PERK was decreased significantly (P<0.05).The results of flow cytometry analysis showed that the apoptotic rates of the HepG 2 cells increased with the increase in SAHA concentration . CONCLUSION:SAHA up-regulates the acetylation of H3K9 and H3K27 in the HepG2 cells and induces apoptosis of HepG2 cells by activating the endoplasmic reticulum stress-related apoptosis pathway .

OBJECTIVE: To analyze the detection rate of the inferior pyloric artery (IPA) in patients with gastric cancer by computed tomography arteriography (CTA). MATERIALS AND METHODS: Fifty-four patients (48 males and 6 females; mean age, 59.0 ± 1.5 years) who had undergone radical gastrectomy for gastric cancer from September 2016 to July 2017 at our institution were recruited prospectively. Patients underwent abdominal contrast-enhanced CT scans and CTA imaging reconstruction before the operation. The origin of the IPA in all cases was determined by a radiologist based on CTA images and verified by the surgeon. The accuracy of CTA in diagnosing the origin of the IPA was calculated. Dominant vessels of the origin were analyzed. RESULTS: IPAs were detected by CTA in 51 patients (94.4%). Among these, IPAs originated from the right gastroepiploic artery (RGEA) (24 cases), the gastroduodenal artery (GDA) (4 cases), and the anterior superior pancreaticoduodenal artery (ASPDA) (20 cases). In the remaining 3 cases, the IPAs contained two branches originating from the RGEA and ASPDA, respectively. During surgery, in 2 (3.7%) of the 54 cases of gastric cancer, IPAs could not be detected; the IPAs originated from the RGEA (22 cases), GDA (5 cases), and ASPDA (24 cases). One case had an IPA originating from both the RGEA and the GDA. Finally, the accuracy of CTA in diagnosing the origin artery of the IPA was 85.2% (46/54). CONCLUSION: CTA can detect the origin of the IPA accurately, which can aid surgeons while performing pylorus-preserving operations.
Angiography ; Arteries ; Female ; Gastrectomy ; Gastroepiploic Artery ; Humans ; Male ; Prospective Studies ; Stomach Neoplasms ; Surgeons ; Tomography, X-Ray Computed